Bioinformatics tools will be generated for a class of ~ 100 proteins known as de- ubiquitinating enzymes (DUBs), which play an important role in cell survival, proliferation, and differentiation. LifeSensor's proprietary SUMO expression enhancement technology permits expression and purification of DUBs with relative ease, and a high throughput microarray of these proteins will afford rapid and cost effective screening. Thus, it is proposed to develop a DUB microarray for the purpose of basic research, biomarker discovery, diagnostics/prognostics, and therapeutics. The DUB microarray will have obvious utility in biomarker discovery and diagnostics development. Expression of DUBs has been associated with diseases such as cancer, diabetes, and Parkinsonism. Over- or under-expression of proteins in disease states tend to elicit elevated or diminished auto-antibody production against those proteins. Thus, auto-antibody profiling can assist in identifying the type and stage of a disease. When a biomarker is identified, it can be further studied for its role in the diagnosis and prognosis of the disease. The particular biomarker could also be a therapeutic target. A DUB microarry not only allows one to perform high throughput screening of samples, but also to evaluate expression levels of DUBs in samples under identical conditions to correlate their effect on each other's expression levels under different circumstances. In the first aim, 10 (ten) DUBs will be immobilized on a variety of solid supports to determine the optimum support in terms of retention of enzyme activity. Next, the catalytic function of the retained DUBs will be compared with their function in a standard assay system. Finally, 50 (fifty) serum samples from the general population will be profiled for variation in the expression of autoantibodies against the DUBs. In Phase II, arrays will be built consisting of all known DUBS for ubiquitin and relevant ubiquitin-like proteins. [unreadable] [unreadable] [unreadable]